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Genotype distribution of hepatitis C virus (HCV) can vary over the years between different patient groups and regions. The prevalence of intravenous drug users (IVDU) is known to increase in our country, yet there are a limited number of studies investigating the distribution of HCV genotypes in this group. These data are essential for monitorization of the changes in HCV epidemiology. The present study aimed to evaluate the five-year results of HCV genotyping among patients infected with HCV related to IVDU and unrelated to drug use. Plasma samples of 720 patients (HCV antibody, HCV RNA positive), which were sent to our laboratory for HCV genotyping between January 2014-March 2019 were analyzed. HCV RNA extraction from plasma samples was performed in the automated-extraction system of EZ1 advanced (Qiagen, Germany) using the EZ1 virus mini kit v2.0 (Qiagen, Germany). Amplicons were obtained by amplifying the 5'NCR and core gene region in the Rotorgene 6000 real-time PCR (Qiagen, Germany) device with the HCV RNA real-time quantitative 2.0 (NLM, Italy) kit. For the genotyping, a commercial line probe assay (LIPA) based on in vitro reverse hybridization GEN-C2.0 kit (NLM, Italy) which can distinguish 1, 2, 3, 4, 6 genotypes and 1a, 1b, 2a/c, 2b, 3a, 3b, 3c, 3k, 4a, 4b, 4c/d, 4e, 4f, 4h, 5a, 6a/b, 6g, 6f/q, 6m, 7a subtypes of HCV, based on variations in the 5'-NCR and core regions was used. HCV genotype distribution of 266 IVDU (93.2%: male; median age: 25 +/- 6.82) and 454 non-drug users (51.3%: male; median age: 56.5 +/- 16.06) were examined. In order of frequency in the group with IVDU; genotype 1a, 3a, 1b, 4c/d, 2b, 4, 3 were observed and genotype 1, 2a/c and mixed genotype (1+3a) were detected in one patient. In the group without IVDU, in order of frequency; genotype 1b, 1a, 3a, 1, 2a/c, 4 were observed and genotype 2b, 4c/d, 5a, 6a/b, 6 and mixed genotype (3+4) were detected in one patient. Genotypes la and 3a were significantly higher in the IVDU group (p< 0.00001, p< 0.00001), while 1b was significantly higher in patients without IVDU (p< 0.00001). Genotypes 1a and 3a were more common in young men (p< 0.00001, p= 0.000163), while 1b was higher in middle-aged women (p< 0.00001). The incidence of genotypes 1b (p= 0.021) and 3a (p= 0.012) was higher in foreign nationals than the Turkish patients. When the HCV genotype distribution was examined by years, it was observed that the percentages of genotype 1b and la were decreasing, while the percentage of genotype 3a was increasing. As a result, in this study, HCV genotype distribution among IVDU was observed to be different from the general population without IVDU. It was found that genotypes la and 3a were more common in the IVDU group. As in the other regions of our country, genotype 1b was found most frequently in the general population. Genotype 3a increases significantly compared to years. In our study, the determination of genotypes existing in different parts of the world may be due to the foreign nationals living in our city and our region is a tourism center. It is also necessary to investigate whether there is an increase in IVDU over the years.

Adenoviruses are wide spectrum illnes apents those we often see as the infections keep in the respiratory system, gastrointestinal system and eyes. The adenoviruses are responsible for the 5-10% of the lower respiratory track infections.In this study the frequency of adenoviruses appearence as lower respiratory track infection in children?s age group is searched by the cell culture, polymerase chain reaction (PCR) and direct fluorescent antibodies staining (DFA) methods and the positivity ratio is found 5,8%. It is found that adenoviruses are especially responsible for bronchitis-bronchiolitis picture but a significiant relationship is not determined between age, accompanying chronic disease, phisical examination, laboratory findings, radiologic findings and seasonal distribution. If cell culture is taken as the reference test PCR and DFA?s sensitivity, specificity, positive predictive value (PPV) and negative predictive volue (PPV) charecteristics are found 100%, 97.5%, 58.3%, 100% and 57%, 95.5%, 40%, 97.7% in order.As a result in this study it is detected that adenoviruses are factors of the cildren?s lower respiratory track infection. However the golden standart for adenoviruses infections diagnosis is the cell culture, PCR method can be used, which is highly sensitive and spesific in diagnosis. DFA?s low sensitivity restricts its application, however it should be kept in mind that the sample quality could be understood by this method. 49

ÖZETSağlık . Balgam Örneklerinde Mikobakterilerin Aranmasında GenoTypeMycobacteria Direct Test Yönteminin Klasik Tanı YöntemleriyleKarşılaştırılması, zole Edilen M. tuberculosis complex'de zoniazid veRifampisin Direncinin Aranması. Eskişehir Osmangazi Üniversitesi TıpFakültesi. Mikrobiyoloji Anabilim Dalı. Tıpta Uzmanlık Tezi. Eskişehir 2007.Tüberkülozun görülme sıklığında son yıllarda gözlenen artış, beraberinde çoklu ilaçdirenci gösteren suşların da yaygınlaşmaları ve bunların önemli sağaltımproblemlerine yol açmaları, etkenin erken tanısı yolundaki arayışları hızlandırmıştır.Mikobakteriyel enfeksiyonların laboratuvar tanısında direkt mikroskopik inceleme vemikroorganizmanın kültürünün yapılması geleneksel yöntemler, GenoTypeMycobacterium Direct test ve MTBDR ise moleküler yöntemlerdir. Çalışmamızda64 balgam örneği; iki farklı direkt mikroskopi yöntemiyle (karbolfuksin ve floresan),iki farklı kültür yöntemiyle (LJ + Bactec 12B) ve GenoType Mycobacteria DirectTest yöntemleriyle incelendiler. Kültürde üreyen örneklerde Bactec 460 TB veGenoType MTBDR ile NH ve R F direnci arandı. Çalışmaya alınan 64 örneğinalındığı hastaların 19'u kadın, 45'i erkekti. Bunların 48'i direkt mikroskopikinceleme ile pozitif olarak değerlendirildi. Bactec 12B'de 45 örnekte üremesaptanırken, LJ besiyerinde 41 örnekte üreme saptandı. GenoType MycobacteriaDirect testinde 50 örnek pozitif olarak değerlendirildi. Kültürde üreyen 45 örnekten4'ünde R F, 3'ünde NH direnci saptandı.BACTEC 460 TB sistemi ve LJ besiyeri kültür kombinasyonu altın standartkabul edilerek floresan boyamanın duyarlılığı %96, özgüllüğü %74, karbolfuksinboyamanın duyarlılığı %80, özgüllüğü %90, tedavi almayan hastalarda GenoTypeMycobacteria Direct testinin duyarlılığı %97, özgüllüğü %100, tedavi alan hastalardaGenoType Mycobacteria Direct testinin duyarlılığı %100, özgüllüğü % 14, GenoTypeMTBDR testinin R F direnci için duyarlılığı % 75, özgüllüğü % 100, NH direnci içinduyarlılığı %60, özgüllüğü ise % 100 olarak hesaplandı.Uygulanan yöntemlerin değerlendirilmesi sonucunda GenoTypeMycobacteria Direct'in ve GenoType MTBDR'nin tüberkülozun hızlı tanısında veilaç direncinin saptanmasında iyi bir potansiyele sahip olduğu sonucuna varılmıştır.Anahtar kelimeler: Balgam, Mycobacteria, tüberküloz, GenoType, direnç ABSTRACTSağlık . Comparison of the Genotype Mycobacteria Direct Assay Method withthe Conventional Identification Methods in the search of Mycobacteria in thesputum samples, search of izoniazid and rifampisin resistance in the M.tuberculosis complex isolated. Eskişehir Osmangazi University, Faculty ofMedicine, Medical Speciality Thesis in Department of Microbiology, Eskişehir,2007. The increase in prevalance of tuberculosis in recent years, wide spread ofmultidrug-resistant strains and important problems in treatment caused by them haveaccelerated the search in rapid diagnosis of the agent. Direct microscopy and cultureof microorganism are conventional methods but GenoType Mycobacteria Direct testand MTBDR are molecular methods in laboratory diagnosis of mycobacteriainfections. In our study, 64 sputum samples were investigated by two different directmicroscopy method (carbolfuchsin and fluorescent), two different culture method (LJ+ Bactec 12B) and GenoType Mycobacteria Direct test. INH and RIF resistance weresearched by Bactec 460 TB in growth samples and by GenoType MTBDR. 19 of 64patients were female and 45 were male. 48 samples were determined positive bydirect microscopy. 45 samples were positive by Bactec 12B, 41 samples werepositive by LJ. 50 samples were positive by GenoType Mycobacteria Direct test. RIFresistance was detected in 4 and INH resistance was detected in 3 of 45 sampleswhich were positive by culture.When we considered the culture combination of Bactec 460 TB system andLJ medium as gold standart, the sensitivity of fluorescent staining was 96%, and thespecificity was 74%, the sensitivity of carbolfuchsin staining was 80%, and thespecificity was 90%. The sensitivity and specificity of GenoType MycobacteriaDirect test, in non-treated patients were 97% and 100%, respectively. In treated-patients the sensitivity and specificity of this test were 100% and 14%, respectively.The sensitivity of GenoType MTBDR test for RIF resistance was 75% and thespecificity was 100%. The sensitivity for INH resistance was 60% and thespecificity was 100%. After evaluation of these methods, we concluded thatGenoType Mycobacteria Direct and GenoType MTBDR have potential in rapiddiagnosis of tuberculosis and determining drug resistance.Key words: Sputum, Mycobacteria, tuberculosis, GenoType, resistance 82